pOET Insect and BacMam Transfer Plasmids

pOET1 Transfer Plasmid Map. pOET1 is a baculovirus transfer plasmid designed for use with the flashBAC™ Baculovirus Expression System and TransIT®-Insect Transfection Reagent. The powerful AcMNPV polyhedrin promoter affords high level gene expression in insect cells. pOET1 is compatible with any baculovirus expression system that utilizes homologous recombination at the polyhedrin locus.

pOET1C_6XHis Transfer Plasmid Map. pOET1C_6XHis is a baculovirus transfer plasmid is designed for use with the flashBAC™ Baculovirus Expression System and TransIT®-Insect Transfection Reagent. The powerful AcMNPV polyhedrin promoter affords high level gene expression in insect cells. A C-terminal 6×His-tag fusion sequence is included for recombinant protein purification with Ni-NTA agarose columns and subsequent removal by thrombin cleavage. pOET1C_6XHis is compatible with any baculovirus expression system that utilizes homologous recombination at the polyhedrin locus.

pOET6 BacMam Transfer Plasmid Map. pOET6 BacMam is a baculovirus transfer plasmid designed for use with the flashBAC™ Baculovirus Expression System and TransIT®-Insect Transfection Reagent. The CMV promoter enables high level gene expression in mammalian cells. pOET6 is compatible with any baculovirus expression system that utilizes homologous recombination at the polyhedrin locus.

flashBAC™ Overview

flashBAC™ DNA Features: flashBAC™ DNA contains a bacterial artificial chromosome (BAC) that enables manufacturing of recombinant virus DNA in E. coli. lef2 and ORF1629 sequences flanking the BAC (in the flashBAC™ DNA) or gene of interest (in the transfer vector) allow for efficient homologous recombination in insect cells. A deletion in ORF1629 prevents replication of parental virus in insect cells. Deletion of the chitinase gene (chiA) maximizes production of secreted proteins and membrane-targeted proteins in insect cells.

Construction of Recombinant Virus Using flashBAC™: For baculovirus production, flashBAC™ DNA is mixed with transfer vector plasmid DNA containing the gene of interest to be inserted into the virus genome. Following co-transfection into insect cells, homologous recombination simultaneously removes the BAC, inserts the gene of interest and and restores ORF1629 allowing the production of infectious virus which can be harvested from the culture medium. Recombinant baculovirus can be used for subsequent transduction of insect or mammalian cells for protein expression.